矮牵牛‘梅林’遗传转化体系的建立  

Establishment of Genetic Transformation System of Petunia hybrida ‘Merlin'

作　　者：曹尚杰 焦孟月 张彦妮[1] 岳莉然[1] CAO Shang-jie;JIAO Meng-yue;ZHANG Yan-ni;YUE Li-ran(College of Landscape Architecture,Northeast Forestry University,Harbin,Heilongjiang 150040,China)

机构地区：[1]东北林业大学园林学院,黑龙江哈尔滨150040

出　　处：《西北林学院学报》2018年第5期123-129,共7页Journal of Northwest Forestry University

基　　金：中央高校基本科研业务费专项资金项目(2572014BA22);黑龙江省留学归国基金项目(LC201410)

摘　　要：在矮牵牛‘梅林’再生体系基础上,建立根癌农杆菌介导的矮牵牛遗传转化体系,以获得转PSARK-IPT基因的矮牵牛植株。结果表明,转化效率最高的预培养时间为2d,农杆菌侵染浓度为OD600=0.5,侵染时间为3min;共培养时间为36h;适宜的抑菌抗生素头孢霉素浓度为500mg·L^(-1);潮霉素作为遗传转化中的筛选标记,选择2mg·L^(-1)为叶片分化筛选压,4mg·L^(-1)的Hey为最佳生根筛选压。对获得的15株潮霉素抗性植株进行PCR及RT-PCR检测,证明7株为阳性,证实目的基因已整合到这7株矮牵牛基因组中。In order to obtain the transgenic plants of the P SARK-IPT gene,Agrobacterium tumefaciens-mediated genetic transformation system was established on the basis of the regeneration system of Petunia hybrida‘Merlin’.The results showed that the best pre-culture time was 2 days,the most suitable A.tumefaciens concentration was OD 600=0.5,the best infection time was 3 minutes,and the best dark culture time was 36 hours.The suitable antibiotic cephalosporin concentration was 500 mg·L-1,2 mg·L-1 Hygromycin as a screening marker in genetic transformation for leaf differentiation screening pressure,and 4 mg·L-1 Hey for the best rooting screening pressure.The 15 plants with hygromycin resistance were detected by PCR and RT-PCR,of which 7 plants were positive.The results confirmed that the target gene had been integrated into the petunia genome.

关 键 词：矮牵牛'梅林’ PSARK-IPT基因 遗传转化 

分 类 号：S681.6[农业科学—观赏园艺]